Automated apparatus and methods for dispensing fluids into microplates utilizing microwell covers

ABSTRACT

Automated apparatus and methods for dispensing fluids into microplates utilizing microwell covers, the covers comprising open portions to allow a pipette access to one or more wells and impermeable portions which prevent the fluids from getting into wells shielded by the impermeable portion. The open portions and impermeable portions are preferably arranged and sized to align with alternating rows of wells in a particular microplate. Preferred covers are movably positioned on the microplate. Automated dispensing apparatus for use with microplates and microwell covers comprises a programmable controller, and suitable interfaces which allow the apparatus to be programmed, and which control a dispensing head such that pipettes are moved in the desired manner in order to take advantage of the protective features of the microwell covers. The apparatus also preferably comprises at least one transfer mechanism for moving a cover relative to a microplate at a dispensing station.

RELATED APPLICATION DATA

The present invention is directed to microwell covers for use withmicroplates used in laboratory testing, automated devices programmed todispense fluid into a microplate using a microwell cover, and methodsfor dispensing fluids into a microplate.

This application is a continuation of U.S. patent application Ser. No.15/646,788 filed on Jul. 11, 2017, which is a continuation of U.S.patent application Ser. No. 14/455,904 filed on Aug. 9, 2014, now U.S.Pat. No. 9,846,172, which is a continuation of U.S. patent applicationSer. No. 13/589,869 filed on Aug. 20, 2012, now U.S. Pat. No. 8,808,644,all of which are hereby incorporated by reference.

BACKGROUND

Microplates, also known as microwell plates and microtitre plates, are astandard product and are regularly used in medical, chemical, andbiological laboratories. Microplates have a plurality of sample wellstypically arranged in a 2:3 rectangular matrix. For example, a commonconfiguration for a microplate has 96 wells arranged in an 8×12 matrix.

In the laboratory, microplates are commonly supplied with variousliquids, e.g. samples, reagents, and solvents. The particular liquidsused will depend on the test being performed. During use, it isimportant to the accuracy of the laboratory procedure being performedthat each liquid be dispensed into the desired well without crosscontamination or unintended dispensing of a liquid into the wrong well.It is also important to the efficient operation of a laboratory to beable to dispense liquids into the desired sample wells accurately andrapidly. Therefore, automated machines have been introduced forautomatically dispensing desired liquids into designated wells of amicroplate. Whether a liquid is dispensed into a sample well or drawnfrom a well, and whether dispensed manually or using an automatedapparatus, the liquid is typically moved with some type of pipette.

Some laboratory procedures utilize two types of microplates, namely,filter plates and collection plates. As these terms are used herein,filter plates have wells, sometimes referred to as columns, withopenings at both the top and bottom, whereas collection plates haveopenings at the top but are closed at the bottom. As used herein, theterm “well” is used to indicate a well of a microplate having either anopen bottom (sometimes referred to as a column) or a closed bottom as ina collection plate. During common procedures, an adsorbent packingmaterial is provided in each well of a filter plate. After the samplefluid has been placed into the well, the adsorbent is washed withsuitable solvents to remove unwanted compounds which are directed to awaste station. Then the filter plate is placed over a collection plateand the desired analyte is removed from the packing material using asuitable eluent and the eluate is collected in the collection plate.

When liquids are moved to any of the wells in a microplate other thanthe wells located on the outer perimeter of the microplate, it isnecessary to move the pipette containing a liquid over wells other thanthe well to which the liquid is intended. This creates a risk ofunintended contamination by a liquid entering a well for which it is notintended.

SUMMARY OF THE INVENTION

Various embodiments of the present invention comprise microwell coversfor microplates. The covers comprise open portions to allow a pipetteaccess to one or more wells and impermeable portions which prevent theliquids from getting into wells shielded by the impermeable portion. Asdescribed in further detail below, preferred embodiments compriseimpermeable portions with receptacles for catching any errant liquidsand open portions with discrete openings sized and configured so that adiscrete opening is provided in the cover for each well in a row of amicroplate. The open portions and impermeable portions are preferablyarranged and sized to align with alternating rows of wells in theparticular microplate with which the cover will be used. Preferredcovers are designed to be movably positioned on the microplate, forexample, by sliding the cover along the top of the microplate in orderto reposition the impermeable portions and open portions of a cover overdifferent wells in the microplate, preferably over different rows ofwells.

Other embodiments comprise automated dispensing apparatus, microplates,and microwell covers. The automated dispensing apparatus comprises aprogrammable controller, such as a microprocessor, and suitableinterfaces, either onboard or external such as a laptop or personalcomputer, which allow the apparatus to be programmed and which control adispensing head such that pipettes are moved in the desired manner inorder to take advantage of the protective features of the microwellcovers. The apparatus also preferably comprises at least one transfermechanism for moving a cover relative to a microplate at a dispensingstation and/or moving microplates and microwell covers from theirrespective supply positions according to preprogrammed instructions.

Other embodiments comprise methods for dispensing fluids to wells in amicroplate. One such method comprises providing a liquid dispenser, amicroplate, and a microwell cover; aligning an open portion of the coverwith at least one row of wells; dispensing at least one liquid into afirst well; subsequently moving the microwell cover relative to themicroplate so that the open portion is aligned with a different row ofwells; and subsequently dispensing at least one liquid into a least onesecond well by passing the liquid through an open portion of themicrowell cover.

Another method comprises providing a liquid dispenser, a microplate, anda microwell cover; moving the dispenser to a position over animpermeable portion of the cover and subsequently to a position over anopen portion; dispensing a liquid into a well by passing liquid throughan open portion of the cover; subsequently moving the dispenser over animpermeable portion; and subsequently moving the dispenser from aposition over an impermeable portion to a position which is not over themicroplate.

These and other aspects are described in further detail below.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a top perspective view of one type of microwell cover of thepresent invention.

FIG. 2 is a bottom perspective view of the cover shown in FIG. 1.

FIG. 3 is a top view of the cover shown in FIG. 1.

FIG. 4 is a right side view of the cover shown in FIG. 1.

FIG. 5 is a front view of the cover shown in FIG. 1.

FIG. 6 is a close up perspective view of a portion of the cover shown inFIG. 1.

FIG. 7 is an exploded perspective view of the cover shown in FIG. 1, afilter plate and a collection plate.

FIG. 8 is a side view of the cover of FIG. 1 on a filter plate in afirst position.

FIG. 9 is a side view of the cover of FIG. 1 on a filter plate in asecond position.

FIGS. 10-18 illustrate one method of the present invention.

FIG. 19 illustrates a microwell cover of an alternative embodiment ofthe present invention.

FIG. 20 illustrates a microwell cover of a third embodiment of thepresent invention.

FIG. 21 illustrates a microwell cover of a fourth embodiment of thepresent invention.

FIG. 22 illustrates a microwell cover of a fifth embodiment of thepresent invention.

FIG. 23 diagrammatically illustrates an automated dispensing apparatusfor performing methods of the present invention.

DETAILED DESCRIPTION

One aspect of the present invention comprises a microplate, a microwellcover configured for use with the microplate, and an automatedlaboratory apparatus comprising liquid dispensing and microplatetransfer mechanisms. The automated laboratory apparatus has the abilityto dispense liquids to at least one and preferably a plurality ofwells/columns in a microplate through one or more dispensing tips. Asused herein, the term “dispensing tips” includes reusable and disposablepipettes, as well as other liquid dispensers suitable for dispensing aliquid into a specific well of a microplate. The dispensing tip isalternatively, releasably attachable or integrally formed with theautomated apparatus. In the case of a disposable pipette, the disposablepipette is releasably connectable to a dispensing head. Preferredembodiments utilize a plurality of releasably attachable dispensing tipsin the form of pipettes, for example, 48 pipettes arranged in 6 rows of8 pipettes in each row.

In use, the cover is positioned on top of the microplate which typicallycomprises a matrix of wells. Commonly used microplates have wellsarranged in a 2:3 rectangular matrix, for example an 8×12 matrixtotaling 96 wells. The present invention is useful with microplateshaving different numbers of wells and different configurations but isillustrated and explained herein with reference to 96 well microplatesfor purposes of illustration.

FIG. 1 is illustrative of one type of microwell cover of the presentinvention designed for use with a 96 well microplate. The illustratedmicrowell cover 10 has a generally rectangular configuration designed torest on a generally rectangular microplate. Cover 10 comprises a frame12, seven rows of receptacles 15 extending generally from the frontportion 11 of the cover to the rear portion 13. The eight receptacles ineach row are configured to align with eight wells in a row of themicroplate. Between each row of receptacles 15 is a row of eightdiscrete openings 30 for a total of six rows of openings. The openings30 in the peripheral row of openings 30 are defined by the frame 12 andboth laterally and longitudinally extending spacers 14. The otheropenings 30 of this embodiment are defined by four laterally andlongitudinally extending spacers 14.

As best shown in the close-up view of FIG. 6, in this preferredembodiment, receptacles 15 are defined by an upwardly extending forwardwall 16, rearward wall 17, left wall 18, right wall 19, and a solidbottom surface 20. Receptacles 15 are designed to catch liquid whichinadvertently drips from a pipette as the pipette is moving to or fromits intended position over a predetermined well of a microplate. Whileother configurations can be used, it is preferable to have the tops ofthe walls separating the receptacles formed as inclined surfaces withoutmuch of a horizontal surface in order to direct any errant droplets intoreceptacle 15. FIG. 6 illustrates an inclined surface 25 formed on theupper regions of each of walls 16-19.

As best illustrated in the bottom view of FIG. 2 and the side view ofFIG. 4, the bottom surface 22 of frame 12 and the downwardly facingsurfaces 24 located below receptacles 15 are generally co-planar in thisillustrated embodiment. It is also feasible to have surfaces 24positioned higher then bottom surfaces 22 of frame 12. In eitherconfiguration it is preferred that the bottom portions of receptacles 15do not extend below the bottom surface 24 of frame 12. This facilitatesthe sliding movement of cover 10 across the top of a microplate asdescribed further below.

Illustrated cover 10 also comprises longitudinally extending front rail40 and rear rail 42 which extend downwardly below the lower surface offrame 12. Rails 40, 42 help to maintain the proper orientation of cover10 when it is positioned on a microplate. Cover 10 also comprises rightand left stops 50 which extend downwardly from frame 12 at the left andright portions of cover 10. Stops 50 are positioned relative to frame 12such that when stops 50 are positioned in abutment with a left or rightedge of a microplate, rows of receptacles 15 and rows of openings 30 arealigned over different rows of wells of a microplate.

FIG. 3 is a top view of microwell cover 10 shown in FIG. 1. For purposesof reference, each column has been numbered 1-13 and each row isdesignated with the letters A-H. FIG. 4 is a right side view of themicrowell shown in FIG. 3, while FIG. 5 is a front view of the cover 10shown in FIG. 3.

From the present descriptions and drawings it will be appreciated thatillustrated cover 10 comprises one more row of total receptacles andopenings than the total number of rows of wells in the illustratedmicroplate. For example, a standard 96 well microplate comprises twelvelaterally extending rows of wells each comprising eight wells.Illustrated cover 10 which is designed for use with a standard 96 wellplate comprises a total of thirteen rows, including seven rows ofreceptacles and six rows of openings. This allows cover 10 to be movedlaterally along the top of a 96 well microplate during a laboratoryprocedure in order to provide access to a first half of the wells whileshielding a second half of the wells when the cover is in one position.After the cover 10 has been moved, the first half of the wells areshielded and the second half are accessible. The wells which are notaccessible at any given time are shielded by receptacles which form animpermeable barrier to errant liquids which might otherwise travel intoan incorrect well. It will be appreciated that frame 12 also providessome protection against airborne droplets which might otherwise enter anincorrect well. According to this illustrated embodiment of the cover,all wells of the microplate are accessible to pipetting during thecourse of a laboratory procedure, but all of the wells are notaccessible at the same time.

FIG. 7 is an exploded view of a microwell cover of the present inventionpositioned above a filter plate 70 which is positioned above acollection plate 75. From the present description those skilled in theart will appreciate that during a collection step, the filter plate 70is positioned on top of the collection plate 75 while the cover 10 ispositioned on top of filter plate 70.

FIGS. 8 and 9 illustrate microwell cover 10 positioned on a deep wellcollection plate 70. In FIG. 8, cover 10 is positioned with left stop 50abutting left edge 72 of the microplate 70 thereby aligning openings 30with the even number rows of wells in microplate 70, namely rows 2, 4,6, 8, 10 and 12. In this position, the receptacles of cover 10 shieldthe wells in the odd numbered rows, namely rows 1, 3, 5, 7, 9 and 11.Thus, when cover 10 is in this position, liquid can be dispensed bydispenser 80 through pipettes 81 into the wells in the accessible evennumbered rows, while reducing the risk of accidentally dripping liquidinto the odd numbered rows.

FIG. 9 illustrates cover 10 positioned with right stop 50 abutting theright edge 73 of microplate 70 in order to align openings 30 with theodd number rows of wells in microplate 70, namely rows 1, 3, 5, 7, 9 and11. Thus it will be appreciated that by moving cover 10 relative tomicroplate 70, half of the wells of the microplate can be madeaccessible to pipettes for the dispensing of fluid or for drawing fluidfrom wells of microplate 70 while the other half of the wells areshielded by the impermeable portions of cover 10 comprising receptacles15.

FIGS. 8 and 9 also illustrate the fingers of a transfer mechanism whichmove covers relative to a microplate, and preferably also transfermicroplates and microwell covers from supply stations to a dispensingstation and then to a disposal station. In FIG. 8, lower fingers 610 and612 hold microplate 70 in a stationary position while left upper finger620 has moved cover 10 to the right until the left stop 50 engages theleft side 72 of microplate 70 in order to properly align the openingsand impermeable portions of cover 10 with the desired wells ofmicroplate 70. In FIG. 9, lower transfer fingers 610 and 612 continue tohold microplate 70 in a stationary position while right upper finger 622has moved cover 10 to the left until right stop 50 has abutted the rightside 73 of microplate 70.

FIGS. 10-18 illustrate advantages of the present invention as well asmethods for distributing liquids to wells in a microplate. Thisembodiment utilizes an automated apparatus comprising a liquid dispenser80 designed to pick up disposable pipettes 81 from a pipette supplyrack, draw a predetermined liquid into each pipette wherein each liquidis to be dispensed into a predetermined specific well, and then to movethe pipettes such that the pipettes do not pass over any open wellsbefore arriving at the well to which the particular liquid in eachpipette is intended.

In FIG. 10 an exemplary liquid dispenser 80 starts from a position whichis not over cover 10 or microplate 70. The dispenser head had previouslybeen moved over a pipette supply rack and lowered into engagement witheight pipettes which are temporarily secured to the dispensing head. Thedispensing head is then moved over at least one liquid supply containerwhere the dispensing head draws a desired liquid into each pipette. Thepipettes can be each supplied with a different liquid, or one or more ofthe pipettes can be supplied with the same liquid. As shown in FIG. 10,the dispensing head is then moved to a position which is higher than andin alignment with an impermeable portion of cover 10 which is blockingliquid access to the wells in row 12. In this position, the dispenser 80is not positioned over either the cover 10 or the microplate 70. InFIGS. 10-18, cover 10 is positioned to the left so that odd numberedrows of wells in microplate 70 are accessible while even numbered rowsof wells are shielded by the impermeable portions of cover 10.

At a desired time, dispenser 80 is moved forwardly in the direction ofarrow F while maintaining the pipettes over receptacles 15 as shown inFIG. 11. With reference to FIG. 12, the dispensing head 80 continues inthe direction of arrow F to move the pipettes 81 until each pipette ispositioned over a receptacle 15 and in lateral alignment with a desiredopening 30 of row 11 in microplate 70. With reference to FIG. 13,dispensing head 80 is then moved to the left in the direction of arrow Lto position each pipette over an opening 30 of row 12 in cover 10 (whichis aligned with the wells in row 11 of the microplate 70). In theposition shown in FIG. 13, the pipettes are disposed directly over wellsin row 11 of microplate 70. If desired, the dispensing head 80 is thenlowered in the direction of arrow D as shown in FIG. 14 causing thepipettes and any liquid contained in the pipettes to move through theopenings of cover 10 and into the desired wells of row 11 where theliquid is then dispensed. With reference to FIG. 15, the dispensing head80 is then raised upwardly in the direction of arrow U to a height whichis higher than the receptacle sidewalls. As shown in FIG. 16, thedispensing head is then moved to the right in the direction of arrow Rin order to position all of the pipettes over receptacles 15. As shownin FIG. 17, the dispensing head 80 is then moved in the direction ofarrow B while keeping the pipettes over receptacles 15 in order to movethe dispensing head 80 to a position where it is no longer positionedover the microplate 70 or cover 10 as shown in FIG. 18.

From the present description it will be appreciated that during thismethod each pipette was never positioned over an unshielded well otherthan the well for which the liquid in that specific well was intended.In this manner, the microwell cover and manner of positioning pipettesover a microplate greatly minimize any chance of contamination from anerrant liquid passing into an unintended well.

It will also be appreciated that the advantages of the present inventionand the minimization of cross contamination are achieved whether thepipettes are dispensing liquid into the wells or drawing liquid fromwells such as the wells of a collection plate. In the case of drawingliquid from wells in a collection plate, the path of the dispensing headis similar except for the elimination of the step of drawing fluid intothe pipettes which is performed when the pipettes are in the positionshown in FIG. 14 rather than when in the position prior to that shown inFIG. 10.

As an alternative to the path of the dispenser 80 shown in FIGS. 10-18,after the dispensing head 80 has dispensed fluid into the open wells,instead of moving the dispensing head to the right (to a position overshielded portions of the cover) and then moving it to a position whichis not over the cover and microplate, after the dispensing head hasdispensed liquid and been raised out of the wells to the position shownin FIG. 15, the head can be moved to the left for a distance of two rowsto dispense fluid into the wells of row nine and can optionally continuein this manner dispensing liquid through each of the open portions intowells in microplate 70.

Dispensing head 80 is shown as supporting a single row of eight pipettesfor clarity of illustration but more preferably, a dispensing head forthe microplate shown in FIGS. 10-18 would comprise six rows of pipetteswith each row having eight pipettes. In such a configuration, all openwells would be supplied with a desired fluid when the pipettes are inthe position illustrated in FIG. 14.

While the configuration of the cover 10 shown in FIGS. 1-7 is presentlypreferred, other configurations of covers may be used. FIG. 19illustrates an alternative cover of the present invention which issimilar to cover 10 shown in FIG. 1 but further comprises an absorbentmaterial 117, such as absorbent paper, cotton, a non-woven material,etc. in each receptacle 115. FIG. 20 illustrates a still furtherembodiment of the present invention which is also similar to theembodiment shown in FIGS. 1-7 however in this embodiment the impermeableportions of cover 210 comprise continuous troughs 215 which extend forthe full length of a row. The side rows defining each trough 215preferably comprise tapered edges similar to tapered edges 25 shown inFIG. 6.

FIG. 21 illustrates a still further embodiment of the present inventioncomprising cover 310. Cover 310 comprises a total of 12 rows of openingsand impermeable portions, i.e. the same number of rows as the microplatefor which cover 310 is intended to be used. In this illustratedembodiment, the impermeable portions comprise strips of absorbent paper315 which are secured between the rows of openings 330. Alternatively,other absorbent or adsorbent materials can be used. According to thisembodiment, rather than simply sliding cover 310 to change the wells towhich a pipette has access, the cover is rotated 180^(N) relative to themicroplate.

FIG. 22 illustrates a still further embodiment of the present inventionwherein both the impermeable portions and portions with openingscomprise walls to separate adjacent portions. The cover 410 shown inFIG. 22 comprises rows of receptacles 415 having closed bottom surfaces420 while adjacent rows of open portions comprise side walls with openbottoms 430. Closed bottoms 420 in each of the impermeable sections,namely the odd numbered rows 1, 3, 5, 7, 9 and 11 are indicated withshading while the even numbered rows have openings to allow liquidaccess to wells aligned with those openings. This embodiment alsocomprises an equal number of rows of openings and impermeablereceptacles but is wide enough for cover 410 to be moved laterally inthe manner illustrated in FIGS. 8 and 9 in order to align openings andshielded areas with different rows of microwells when desired.

The covers are preferably formed of materials which will have sufficientdurability during their expected life span in the environments in whichthey will be used, including contact with the fluids used during variousprocedures. For example, suitable materials for most applicationsinclude polyethylene, polypropylene and Teflon®.

While the covers of the present invention and the methods described canbe utilized when liquid is manually dispensed into a microplate, it ispreferable to use an automated laboratory apparatus to increase speed,accuracy and efficiency of a laboratory processing multiple samples.FIG. 23 diagrammatically illustrates a automatic laboratory apparatus600 comprising a housing 605 and a dispensing head 680 supported by adispensing arm 685 which is movable horizontally, vertically, and intoand out of housing 605 as indicated by the arrows in FIG. 23. Apparatus600 also comprises a transfer mechanism comprising lower fingers 610,612 and upper fingers 620, 622. Each of the fingers of the transfermechanism can preferably be moved independently of each other as well ashorizontally, vertically and into and out of housing 605. A programmablecontroller 500, linked to a suitable input device 510 such as a computeror touch screen, is used to control the operation and movement ofdispensing head 680 and fingers 610, 612, 620 and 622.

This illustrated embodiment comprises a dispensing station 630, apipette supply station 640, a liquid supply station 650, a pipettedisposal area 645, a filter plate supply 660, a collection plate supply670 and a microwell cover supply 675.

Controller 500 is programmable to cause transfer mechanism, preferablylower fingers 610 and 612 to initially move a filter plate from filterplate supply 670 to dispensing station 630, then a cover from coversupply 675 is retrieved by upper fingers 620, 622 and positioned on topof a filter plate at the dispensing station 630. Simultaneously orsequentially with the transfer of the filter plate and cover todispensing station 630, dispensing head 680 picks up pipettes frompipette supply 640 and draws the desired liquid into those pipettes atliquid supply station 650. Dispensing head 680 then moves the pipettesto the desired wells, preferably in the manner described above withrespect to FIGS. 10-18. After liquid has been dispensed into some of thewells in a microplate, the cover is moved relative to the microplate bythe transfer fingers in the manner illustrated above in FIGS. 8 and 9.Namely, lower fingers 610, 612 can grasp the filter plate and hold it inplace while at least one of upper fingers 620, 622 moves the coverrelative to the microplate in order to align the openings of the coverwith different columns in the microplate. Alternatively, the cover iskept in place and the plate is moved, or both are moved in oppositedirections. After liquid has been dispensed as desired and dispensinghead 680 is moved to a position where it is no longer positioned overdispensing station 630, the pipettes are ejected into pipette disposalcontainer 645. When it is desired to collect sample from a filter plate,transfer arms can be used to position a collection plate under thefilter plate.

Thus the automated laboratory apparatus is preferably designed toretrieve microplates and covers from respective supply racks, positionthe cover on a microplate in a desired position, move the cover relativeto the microplate when desired and remove the cover from the microplateafter the desired dispensing and/or withdrawing of fluids has occurred.The automated apparatus also moves each dispensing tip, e.g. pipette, ina manner such that the dispensing tips are moved over shielded portionsuntil arriving at the opening aligned with the well to which the liquidin a given dispensing tip is intended to be dispensed.

The invention claimed is:
 1. A method for dispensing fluid into amicroplate comprising the steps of: providing a microplate comprising aplurality of wells; providing a microwell cover comprising: at least oneimpermeable portion configured to shield a first plurality of wells; andat least two open portions wherein a first open portion is on a firstside of said impermeable portion and a second open portion is on asecond side of said impermeable portion which is different from saidfirst side, said open portions are configured to be aligned above asecond plurality of wells while permitting access to wells in the secondplurality of wells; movably positioning said microwell cover on saidmicroplate; aligning at least one of said at least one open portion withat least one first well in a first row; dispensing a fluid into at leastone first well by passing said fluid through said at least one openportion; subsequently moving said microwell cover relative to saidmicroplate so that said open portion is aligned with a different well;and subsequently dispensing at least one fluid into at least one secondwell by passing said fluid through an open portion of a said microwellcover, wherein said second well is different from said first well.